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Live Imaging Assay for Assessing the Roles of Ca2+ and Sphingomyelinase in the Repair of Pore-forming Toxin Wounds
Published on: August 25, 2013
High-Throughput Mechanical Rupture of Nuclear Envelope and the Intracellular Dynamics of Massive Wound Repair
Apresio K Fajrial1, Leyla Akh2, Stephanie E Schneider1
1Department of Mechanical Engineering, University of Colorado, Boulder, Colorado, USA.
Abstract:
The dynamics of nuclear envelope rupture and wound repair are critical biological processes that play essential roles in cell homeostasis. Previous studies on nuclear envelope repair dynamics have mainly focused on small ruptures induced by low-throughput tools such as laser ablation and atomic force microscopy. Here, a device is presented that deterministically porates the cell membrane and nuclear envelope in high throughput, with applications in single-cell studies of wound repair dynamics and in statistical assessments of cell poration. The device consists of sharp nanostructures in arrays of microchannels, enabling precise and localized disruption at both the plasma membrane and nuclear envelope while preserving cell viability. The distribution of endosomal sorting complexes required for transport (ESCRT) proteins after mechanical disruption at both the plasma membrane and nuclear envelope highlights the cell recovery strategy for severe wounds. Interestingly, during this extreme rupture, resources seem to be allocated more toward nuclear envelope repair. High-throughput mechanoporation on membrane systems provides a new perspective on studying complex wound repair dynamics, opening promising avenues for further research in the field of wound healing mechanisms.
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